Yoshimura F K, Yamamura J M
J Virol. 1981 Jun;38(3):895-905. doi: 10.1128/JVI.38.3.895-905.1981.
Four cloned rat cell lines (NX-1 to -4) infected with Moloney murine leukemia virus and defective in virus replication were found to be all different by viral protein and nucleic acid analyses. All four clones produced noninfectious particles and, except for NX-2, at about the same level as wild type. Compared with wild-type virions these defective particles contained larger amounts of gag precursor proteins and very little or no p30 or p15. Analysis of intracellular precursor proteins revealed that NX-2 to -4 synthesized normal Pr65gag, whereas NX-1 produced a slightly smaller precursor. Both NX-1 and NX-4 synthesized an intracellular polyprotein with a size similar to that of wild-type Pr180 gag-pol. Restriction endonuclease analysis of NX-1 to -4 cellular DNA showed that each clone contained a single integrated provirus which possessed large terminal repeat sequences at both the 5' and 3' ends. The proviruses of NX-1 to -3 appeared normal by restriction endonuclease analysis, but NX-4 provirus had a deletion of 1,700 base pairs comprising part of the polymerase region. The noninfectious particles produced by all four clones packaged Moloney viral RNAs and rat RNAs of two different sizes.
通过病毒蛋白和核酸分析发现,四种感染莫洛尼氏鼠白血病病毒且病毒复制存在缺陷的克隆大鼠细胞系(NX-1至-4)各不相同。所有四个克隆均产生无感染性的颗粒,除NX-2外,其产生水平与野生型大致相同。与野生型病毒粒子相比,这些缺陷颗粒含有大量的gag前体蛋白,而p30或p15含量极少或没有。细胞内前体蛋白分析表明,NX-2至-4合成正常的Pr65gag,而NX-1产生的前体略小。NX-1和NX-4均合成了一种大小与野生型Pr180 gag-pol相似的细胞内多蛋白。对NX-1至-4细胞DNA进行限制性内切酶分析表明,每个克隆均含有一个单一整合的前病毒,其5'和3'末端均具有大的末端重复序列。通过限制性内切酶分析,NX-1至-3的前病毒似乎正常,但NX-4前病毒缺失了1700个碱基对,该缺失部分包括聚合酶区域。所有四个克隆产生的无感染性颗粒包装了莫洛尼氏病毒RNA和两种不同大小的大鼠RNA。